In addition, the analysis demonstrated the presence of constituents that are usually not considered in the standardization protocols (shikimic acid and 3,4-dihydroxy benzoic acid) as well as excipients (glycerol and citric acid). of components. Electronic supplementary material The online version of this article (doi:10.1007/s11306-009-0195-x) contains supplementary material, which is available to authorized users. leaves are among the best-selling natural preparations worldwide (Warrier and Corzine 2000). Originally, only the seeds were used in traditional Chinese medicine (Bensky et al. 2004), but in the 1960s it was discovered that leaf components improved central and peripheral blood circulation (DeFeudis 1991; Drieu and Jaggy 2000). This led to the development of standardized leaf components (named EGb 761) comprising 6% of TTLs (3.1% of ginkgolides and 2.9% of bilobalide) and 24% of flavonol glycosides (Drieu and Jaggy 2000). Since then, the interest in crude Ceftaroline fosamil acetate as well as standardized components offers increased dramatically, and a series of excellent reviews of the chemistry and biology (Str?mgaard and Nakanishi 2004; Singh et al. 2008), pharmacology (Maclennan et al. 2002; Ahlemeyer and Krieglstein 2003; Schulz 2003), analytical methods (vehicle Beek 2002; vehicle Beek and Montoro 2009), and chromatographic and spectroscopic properties (vehicle Beek 2005) have been published. The primary constituents of standardized leaf components are flavonol glycosides displayed by constructions 1C8 identified with this work, ginkgolides A (GA, 9), B (GB, 10), C (GC, 11), and J (GJ, 12), and bilobalide (BB, 13) (Fig.?1). Additional major classes of compounds (content material 5%) found in the standardized draw out are proanthocyanidins, carboxylic acids, and non-flavonoid glycosides, whereas biflavones and alkylphenols (ginkgolic acids, ginkgols and bilobols) are eliminated during the developing process (vehicle Beek and Montoro 2009). The standardized components are amongst additional utilized for symptomatic treatment of dementia, Alzheimers disease, peripheral occlusive arterial disease, and tinnitus (Mahady 2001), and both the TTLs and flavonoid glycosides are thought to contribute to the neuroprotective effect. Therefore, in 1985 it was discovered that Ceftaroline fosamil acetate ginkgolides are antagonists of the platelet-activating element receptor, which is definitely involved in slowing the progression of neurodegenerative diseases (Singh and Saraf 2001). Recently it was found that GB is an antagonist of the glycine receptors and BB is an antagonist of the -aminobutyric acid receptors (Ivic et al. 2003). Flavonoid glycosides are antioxidants that can potentially prevent neurodegenerative Ceftaroline fosamil acetate diseases caused by oxidative stress (Ramassamy 2006), and quercetin offers been shown to enhance serotonin uptake in synaptosomes from mouse cortex (Ramassamy et al. 1992). Several animal studies and clinical tests support the effectiveness of the standardized draw out, but the precise mechanism and the constituents responsible Ceftaroline fosamil acetate for the effect remain largely unknown due to contradictory results. One reason for this could be that the majority of investigations are based on components that are standardized using methods, which do not assure the same batch-to-batch or brand-to-brand distribution of individual TTLs and flavonoid glycosides. In addition, additional constituents than TTLs and flavonoid glycosides may contribute to pharmacological activity without being included in the standardization. Open in a separate windowpane Fig.?1 Structure of flavonoid glycosides 1C8 and terpene trilactones 9C13 recognized in commercially available preparations To ameliorate the above problems, there is a need for a non-selective analytical technique that allows assessment of the global composition of the extract. Such a method should be complementary to the existing targeted methods based on HPLC coupled with evaporative light-scattering detector for analysis of TTLs SPRY4 (Ganzera et al. 2001), with UVCVIS or PDA for analysis of flavonoid glycosides (Hasler et al. 1992b), and with various types of MS for independent or simultaneous detection of TTLs and flavonoids (Li et al. 2002; Sun et al. 2005; Ding et al. 2006). High-field 1H NMR spectroscopy can, in one spectrum acquired within a few minutes, provide a non-selective metabolic fingerprint of all hydrogen-containing organic constituents in an draw out present above the detection threshold. Because of this, 1H NMR-based metabolomics (Nicholson et al. 2007) offers proven important for data-driven analysis of complex mixtures like natural preparations and medicinal vegetation, including (Rasmussen et Ceftaroline fosamil acetate al. 2006; Lawaetz et al. 2009), (Bailey et al. 2002), varieties (Kim et al. 2005), varieties (Frdrich et al. 2004), (Wang et al. 2004), and (Choi et al. 2004). In the current work, 1H NMR-based metabolomics was utilized for investigation of the global composition of 16 commercially available preparations, and HPLC-PDA-MS-SPE-NMR (Staerk et al. 2006) was utilized for unambiguous recognition of eight major flavonoid glycosides. Materials and methods General experimental methods 1H NMR spectra of components of preparations were recorded at 25C using a Bruker Avance spectrometer (1H resonance rate of recurrence 600.13?MHz) equipped with a 5?mm 1H13C probe. HPLC-PDA-MS-SPE-NMR experiments were performed on a system consisting of an Agilent 1100 LC system, a Knauer K100.